title: Transcriptional regulation of claudin-18 via specific protein kinase C signaling pathways and modification of DNA methylation in human pancreatic cancer cells.
abstract: Since claudin-18 (Cldn18) is overexpressed in precursor lesion PanIN and pancreatic duct carcinoma, it serves as a diagnostic marker and a target of immunotherapy. The stomach isoform of Cldn18, Cldn18a2 is regulated via a PKC/MAPK/AP-1-dependent pathway in PKC activator 12-O-tetradecanoylphorbol 13-acetate (TPA)-stimulated gastric cancer cells. However, little is known about how Cldn18 is regulated, not only in pancreatic duct carcinoma but also in normal human pancreatic duct epithelial cells (HPDE cells). In the present study, four pancreatic cancer cell lines, HPAF-II, HPAC, PANC-1 and BXPC3, and hTERT-HPDE cells in which the hTERT gene was introduced into HPDE cells in primary culture, were treated with TPA. In all human pancreatic cancer cell lines and hTERT-HPDE cells, Cldn18 mRNA indicated as Cldn18a2 was markedly induced by TPA and in well- or moderately differentiated human pancreatic cancer cells HPAF-II and HPAC and hTERT-HPDE cells, the protein was also strongly increased. The upregulation of Cldn18 by TPA in human pancreatic cancer cell lines was prevented by inhibitors of PKCδ, PKCε, and PKCα, whereas the upregulation of Cldn18 by TPA in hTERT-HPDE cells was prevented by inhibitors of PKCδ, PKCθ, and PKCα. Furthermore, a CpG island was identified within the coding sequence of the Cldn18 gene and treatment with the demethylating agent 5-azadeoxycytidine enhanced upregulation of Cldn18 by TPA in HPAF-II and HPAC, but not hTERT-HPDE cells. Our findings suggest that in human pancreatic cancer cells, Cldn18 is primarily regulated at the transcriptional level via specific PKC signaling pathways and modified by DNA methylation.
authors.affiliation: Department of Surgery, Sapporo Medical University School of Medicine, Sapporo, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Ito
authors.forename: Takashi
authors.initials: T
authors.lastname: Kojima
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yamaguchi
authors.forename: Daisuke
authors.initials: D
authors.lastname: Kyuno
authors.forename: Yasutoshi
authors.initials: Y
authors.lastname: Kimura
authors.forename: Masafumi
authors.initials: M
authors.lastname: Imamura
authors.forename: Akira
authors.initials: A
authors.lastname: Takasawa
authors.forename: Masaki
authors.initials: M
authors.lastname: Murata
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tanaka
authors.forename: Koichi
authors.initials: K
authors.lastname: Hirata
authors.forename: Norimasa
authors.initials: N
authors.lastname: Sawada
mesh_terms: D021441:Carcinoma, Pancreatic Ductal / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D057167:Claudins; D019175:DNA Methylation; D004789:Enzyme Activation; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000378:metabolism; D008301:Maleimides / Q000494:pharmacology; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D010183:Pancreatic Ducts / Q000378:metabolism / Q000473:pathology; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D019108:Tight Junctions / Q000378:metabolism; D015854:Up-Regulation
chemical_list: C486683:CLDN18 protein, human; D057167:Claudins; D007211:Indoles; D007527:Isoenzymes; D008301:Maleimides; D008565:Membrane Proteins; D011493:Protein Kinase C; C070515:bisindolylmaleimide I; D013755:Tetradecanoylphorbol Acetate